Dynamic Vascular Developmental Model Of Mirna Changes

Objective To establish chick embryo chorioallantoic membrane dynamic vascular development model, observe and analyse the change of miRNA expression in the process of vascular development. To determine vascular development and key regulatory miRNA and its downstream genes provide experimental basis.Methods On basis of chicken chorioallantoic membrane vascular short development cycle and good maneuverability of the technique, select fresh fertile eggs and incubation. In situ observe the vascular development of chick embryo chorioallantoic membrane from the first day to the 21st day and identify the three time points of chorioallantoic blood vessel: neovascularization, exuberant and obliterated, establish dynamic vascular development model. Select chorioallantoic membrane vascular extract protein of different time points , Detect FGF2 and TGF-β2 protein in chorioallantoic membrane of this three-stage by Western blotting, judge whether to establish a model of dynamic vascular development successful. Extract total RNA by Trizol, UV absorption assay measuring RNA concentration and purity, 28s,18s bands were detected in agarose gel electrophoresis, the quality of RNA was checked, produce miRNA array, analyse miRNA changes in the process of vascular development.Results Research found that the 4 day, 13 days, 16 days respectively for the three time points according to the development of vascular neovascularization, exuberant and obliterated; westen blot results consistent with the literature; miRNA expression changes in vascular development process:⑴, brisk period compared with the newborn : 5 miRNA increased expressed, 5 miRNA reduced expressed;⑵. dissipated period compared with the newborn : upregulation of 37 miRNA, downregulation of four miRNA;⑶, dissipated period compared with the exuberant period: upregulation of 27 miRNA, down regulation of two miRNA.Conclusions1. On basis of chicken chorioallantoic membrane vascular development cycle is short and repeatable characteristics, the newborn period, the exuberant period, Different stages dissipated in the process of vascular development are easily distinguished, we can establish a new dynamic model of vascular development.2. For vascular development of the newborn period, the exuberant period dissipated period in different phases corresponding to the miRNA expression profiles of notable changes , we provide to identify key vascular development and downstream control miRNA genes with new ideas and direction.